Background

Humanized mouse models are still under development, and various protocols exist to improve human cell engraftment and function.

Methods

Fourteen NOD/SCID/IL-2Rγnull (NSG) mice (4‒5 wk old) were conditioned with busulfan and injected with human umbilical cord blood (hUCB)-derived CD34⁺ hematopoietic stem cells (HSC) via retro-orbital sinuses. The bone marrow (BM), spleen, and peripheral blood (PB) were analyzed 8 and 12 weeks after HSC transplantation.

Results

Most of the NSG mice tolerated the regimen well. The percentage of hCD45⁺ and CD19⁺ cells rose significantly in a time-dependent manner. The median percentage of hCD45⁺cells in the BM was 55.5% at week 8, and 67.2% at week 12. The median percentage of hCD45⁺ cells in the spleen at weeks 8 and 12 was 42% and 51%, respectively. The median percentage of hCD19⁺ cells in BM at weeks 8 and 12 was 21.5% and 39%, respectively (P=0.04). Similarly, the median percentage of hCD19⁺ cells in the spleen at weeks 8 and 12 was 10% and 24%, respectively (P=0.04). The percentage of hCD19⁺ B cells in PB was 23% at week 12. At week 8, hCD3⁺ T cells were barely detectable, while hCD7⁺ was detected in the BM and spleen. The percentage of hCD3⁺ T cells was 2‒3% at week 12 in the BM, spleen, and PB of humanized NSG mice.

Conclusion

We adopted a simplified protocol for establishing humanized NSG mice. We observed a higher engraftment rate of human CD45⁺ cells than earlier studies without any significant toxicity. And human CD45⁺ cell engraftment at week 8 was comparable to that of week 12.

Keywords: Humanized mice, Busulfan, Retro-orbital sinus, Hematopoietic stem cell